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On-line Access: 2016-06-03

Received: 2015-10-31

Revision Accepted: 2016-01-22

Crosschecked: 2016-05-12

Cited: 2

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Citations:  Bibtex RefMan EndNote GB/T7714

 ORCID:

Feng-hua Liu

http://orcid.org/0000-0002-6428-4052

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Journal of Zhejiang University SCIENCE B 2016 Vol.17 No.6 P.413-424

http://doi.org/10.1631/jzus.B1500261


Magnolol pretreatment attenuates heat stress-induced IEC-6 cell injury


Author(s):  Chen Mei, Sha-sha He, Peng Yin, Lei Xu, Ya-ran Shi, Xiao-hong Yu, An Lyu, Feng-hua Liu, Lin-shu Jiang

Affiliation(s):  Beijing Key Laboratory for Dairy Cow Nutrition, College of Animal Science and Technology, Beijing University of Agriculture, Beijing 102206, China; more

Corresponding email(s):   liufenghua1209@126.com, jls@bac.edu.cn

Key Words:  Cell-cycle arrest, Cell injury, Heat stress, IEC-6 cell line, Magnolol


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Chen Mei, Sha-sha He, Peng Yin, Lei Xu, Ya-ran Shi, Xiao-hong Yu, An Lyu, Feng-hua Liu, Lin-shu Jiang. Magnolol pretreatment attenuates heat stress-induced IEC-6 cell injury[J]. Journal of Zhejiang University Science B, 2016, 17(6): 413-424.

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author="Chen Mei, Sha-sha He, Peng Yin, Lei Xu, Ya-ran Shi, Xiao-hong Yu, An Lyu, Feng-hua Liu, Lin-shu Jiang",
journal="Journal of Zhejiang University Science B",
volume="17",
number="6",
pages="413-424",
year="2016",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B1500261"
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%T Magnolol pretreatment attenuates heat stress-induced IEC-6 cell injury
%A Chen Mei
%A Sha-sha He
%A Peng Yin
%A Lei Xu
%A Ya-ran Shi
%A Xiao-hong Yu
%A An Lyu
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A1 - Peng Yin
A1 - Lei Xu
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A1 - Xiao-hong Yu
A1 - An Lyu
A1 - Feng-hua Liu
A1 - Lin-shu Jiang
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DOI - 10.1631/jzus.B1500261


Abstract: 
Objective: heat stress (HS) is an important environmental stressor that adversely influences livestock during the summer. The aim of this study was to investigate whether magnolol protects against HS-induced intestinal epithelial cell injury. Materials and methods: An intestinal epithelial cell line (IEC-6) was subjected to HS at 42 °C, with and without magnolol pretreatment. cell injury was detected by monitoring lactate dehydrogenase (LDH) release. MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) assay was used to assess cell proliferation and viability, including identifying effective concentrations of magnolol. Flow cytometry confirmed G1-phase cell-cycle arrest and its alleviation by magnolol. Active DNA synthesis was measured by incorporation of nucleic acid 5-ethynyl-2'-deoxyuridine (EdU). G1-phase cell-cycle-related gene expression was assessed by real-time reverse transcription polymerase chain reaction (RT-PCR) and levels of G1-phase-related proteins by Western blotting. Results: HS induced IEC-6 cell injury and decreased cell viability, as demonstrated by data from LDH and MTS assays, respectively. Based on a number of criteria, IEC-6 cells subjected to HS were arrested in the G1 phase of the cell cycle. magnolol pretreatment decreased HS-induced cell injury through relief of this cell-cycle arrest. Conclusions: magnolol pretreatment attenuates HS-induced injury in IEC-6 cells. magnolol is potentially promising as a protective strategy for HS in livestock.

厚朴酚缓解热应激诱导的小肠上皮细胞损伤及其分子机制

目的:探讨厚朴酚(Mag)对热应激大鼠小肠上皮细胞(IEC-6)修复作用,并初步探讨其作用机制。
创新点:首次在大鼠小肠上皮细胞热应激模型中证明厚朴酚可明显降低细胞损伤程度,且此作用与细胞周期G1期相关。
方法:本试验采用大鼠小肠上皮细胞(IEC-6)为研究对象,将其分为五组:对照组(37 °C,5% CO2),热应激组(42 °C,3 h),热应激+厚朴酚低浓度组(5 µmol/L),热应激+厚朴酚中浓度组(10 µmol/L),热应激+厚朴酚高浓度组(20 µmol/L)。采用MTS法复制热应激模型及厚朴酚药物浓度筛选;采用流式细胞术检测热应激造成大鼠小肠上皮细胞细胞周期阻滞及不同浓度厚朴酚缓解细胞周期阻滞情况;利用电子显微镜观察热应激造成的细胞损伤情况;利用荧光电子显微镜观察EdU染色后,热应激对细胞增殖情况的影响及不同浓度厚朴酚的修复作用;采用荧光定量聚合酶链反应(PCR)技术检测热应激对细胞周期基因表达影响及不同浓度厚朴酚对细胞周期基因调节作用,采用蛋白质免疫印迹(Western blot)技术检测热应激对细胞周期蛋白表达的影响及不同浓度厚朴酚对细胞周期蛋白修复作用。
结论:流式细胞术显示热应激造成IEC-6细胞周期阻滞在G1期(96.5%),而三种浓度厚朴酚均可以缓解细胞周期阻滞现象且呈剂量依赖性(5 µmol/L 88.8%,10 µmol/L 81.0%,20 µmol/L 73.5%)。热应激导致阻滞细胞分裂的G1期基因p21p27Rb显著上调(P<0.01),显示细胞周期阻滞,不同浓度厚朴酚下调这三个基因表达且呈剂量依赖性;促进细胞分裂的G1期基因E2F1CDK4cyclin D1表达显著下调(P<0.01),显示细胞周期阻滞不能正常增值,厚朴酚有效上调这三个基因表达水平。热应激导致G1期阻滞细胞分裂蛋白p21及p27上调显著(P<0.01),厚朴酚有效下调p21及p27蛋白表达,且在20 µmol/L浓度时效果最佳;而G1期促进细胞分裂蛋白pRb、E2F1、CDK4和cyclin D1(CCND1)在热应激下表达显著下调(P<0.01),厚朴酚具有一定上调以上四个蛋白的作用,且在20 µmol/L浓度时效果最佳。厚朴酚作为天然成分药物,通过缓解热应激造成的IEC-6细胞G1期细胞周期阻滞,具有缓解热应激造成的细胞损伤能力,有望作为预防畜禽热应激的饲料添加剂。

关键词:细胞周期阻滞;细胞损伤;热应激;IEC-6;厚朴酚

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Reference

[1]Adnane, J., Shao, Z., Robbins, P.D., 1999. Cyclin D1 associates with the TBP-associated factor TAFII250 to regulate Sp1-mediated transcription. Oncogene, 18(1):239-247.

[2]Alekseenko, L.L., Zemelko, V.I., Domnina, A.P., et al., 2014. Sublethal heat shock induces premature senescence rather than apoptosis in human mesenchymal stem cells. Cell Stress Chaperon., 19(3):355-366.

[3]Bang, J., Huh, J.H., Na, J.W., et al., 2015. Cell proliferation and motility are inhibited by G1 phase arrest in 15-kDa selenoprotein-deficient Chang liver cells. Mol. Cells, 38(5):457-465.

[4]Chang, C.P., Hsu, Y.C., Lin, M.T., 2003. Magnolol protects against cerebral ischaemic injury of rat heatstroke. Clin. Exp. Pharmacol. Physiol., 30(5-6):387-392.

[5]Chen, S.C., Chang, Y.L., Wang, D.L., et al., 2006. Herbal remedy magnolol suppresses IL-6-induced STAT3 activation and gene expression in endothelial cells. Br. J. Pharmacol., 148(2):226-232.

[6]Coqueret, O., 2002. Linking cyclins to transcriptional control. Gene, 299(1-2):35-55.

[7]de Maria, S., Scognamiglio, I., Lombardi, A., et al., 2013. Polydatin, a natural precursor of resveratrol, induces cell cycle arrest and differentiation of human colorectal Caco-2 cell. J. Transl. Med., 11(1):264.

[8]Fofaria, N.M., Kim, S.H., Srivastava, S.K., 2014. Piperine causes G1 phase cell cycle arrest and apoptosis in melanoma cells through checkpoint kinase-1 activation. PLoS ONE, 9(5):e94298.

[9]Gao, C.C., Ma, T.H., Pang, L.Q., et al., 2014. Activation of p21-activated protein kinase 2 is an independent prognostic predictor for patients with gastric cancer. Diagn. Pathol., 9(1):55.

[10]Gao, C.Q., Zhao, Y.L., Li, H.C., et al., 2015. Heat stress inhibits proliferation, promotes growth, and induces apoptosis in cultured Lantang swine skeletal muscle satellite cells. J. Zhejiang Univ.-Sci. B (Biomed. & Biotechnol.), 16(6):549-559.

[11]Guerra-Araiza, C., Alvarez-Mejia, A.L., Sanchez-Torres, S., et al., 2013. Effect of natural exogenous antioxidants on aging and on neurodegenerative diseases. Free Radic. Res., 47(6-7):451-462.

[12]Guyot, S., Gervais, P., Young, M., et al., 2015. Surviving the heat: heterogeneity of response in Saccharomyces cerevisiae provides insight into thermal damage to the membrane. Environ. Microbiol., 17(8):2982-2992.

[13]Intasqui, P., Antoniassi, M.P., Camargo, M., et al., 2015. Differences in the seminal plasma proteome are associated with oxidative stress levels in men with normal semen parameters. Fertil. Steril., 104(2):292-301.

[14]Kaija, H., Pakanen, L., Kortelainen, M.L., et al., 2015. Hypothermia and rewarming induce gene expression and multiplication of cells in healthy rat prostate tissue. PLoS ONE, 10(5):e0127854.

[15]Khosravinia, H., 2015. Effects of Satureja khuzistanica essential oils in drinking water on mortality, production performance, water intake, and organ weights in broiler chickens reared under heat stress condition. Int. J. Biometeorol., 59(11):1711-1719.

[16]Kong, C.W., Tsai, K., Chin, J.H., et al., 2000. Magnolol attenuates peroxidative damage and improves survival of rats with sepsis. Shock, 13(1):24-28.

[17]Kuhl, N.M., Rensing, L., 2000. Heat shock effects on cell cycle progression. Cell Mol. Life Sci., 57(3):450-463.

[18]Lee, K.L., Niu, K.C., Lin, M.T., et al., 2013. Attenuating brain inflammation, ischemia, and oxidative damage by hyperbaric oxygen in diabetic rats after heat stroke. J. Formos. Med. Assoc., 112(8):454-462.

[19]Lei, L., Hepeng, L., Xianlei, L., et al., 2013. Effects of acute heat stress on gene expression of brain-gut neuropeptides in broiler chickens. J. Anim. Sci., 91(11):5194-5201.

[20]Loong, C.C., Chiu, J.H., Tiao, R.C., et al., 2002. Protective effect of magnolol on the small intestinal ischemia-reperfusion injury. Transplant. Proc., 34(7):2679-2680.

[21]McMahon, C., Suthiphongchai, T., Direnzo, J., et al., 1999. P/CAF associates with cyclin D1 and potentiates its activation of the estrogen receptor. PNAS, 96(10):5382-5387.

[22]Men, X., Wang, L., Yu, W., et al., 2015. Cullin7 is required for lung cancer cell proliferation and is overexpressed in lung cancer. Oncol. Res., 22(2):123-128.

[23]Min, L., Cheng, J.B., Shi, B.L., et al., 2015. Effects of heat stress on serum insulin, adipokines, AMP-activated protein kinase, and heat shock signal molecules in dairy cows. J. Zhejiang Univ.-Sci. B (Biomed. & Biotechnol.), 16(6):541-548.

[24]Miova, B., Dinevska-Kjovkarovska, S., Esplugues, J.V., et al., 2015. Heat stress induces extended plateau of Hsp70 accumulation—a possible cytoprotection mechanism in hepatic cells. J. Cell. Biochem., 116(10):2365-2374.

[25]Mitrea, D.M., Yoon, M.K., Ou, L., et al., 2012. Disorder-function relationships for the cell cycle regulatory proteins p21 and p27. Biol. Chem., 393(4):259-274.

[26]Pan, Z., Shao, Y., Dong, W., et al., 2014. Xuebijing attenuates hypotension through the upregulation of angiotensin II type 1 receptor-associated protein 1 in rats suffering from heat stroke. Int. J. Mol. Med., 34(6):1699-1705.

[27]Qi, D., Hu, Y., Li, J., et al., 2015. Hyperthermia induces apoptosis of 786-O cells through suppressing Ku80 expression. PLoS ONE, 10(4):e0122977.

[28]Rasul, A., Yu, B., Khan, M., et al., 2012. Magnolol, a natural compound, induces apoptosis of SGC-7901 human gastric adenocarcinoma cells via the mitochondrial and PI3K/AKT signaling pathways. Int. J. Oncol., 40(4):1153-1161.

[29]Sarkar, R., Mukherjee, A., Mukherjee, S., et al., 2014. Curcumin augments the efficacy of antitumor drugs used in leukemia by modulation of heat shock proteins via HDAC6. J. Environ. Pathol. Toxicol. Oncol., 33(3):247-263.

[30]Song, X., Luo, J., Fu, D., et al., 2014. Traditional Chinese medicine prescriptions enhance growth performance of heat stressed beef cattle by relieving heat stress responses and increasing apparent nutrient digestibility. Asian-Australas J. Anim. Sci., 27(10):1513-1520.

[31]Song, Z., Liu, L., Sheikhahmadi, A., et al., 2012. Effect of heat exposure on gene expression of feed intake regulatory peptides in laying hens. J. Biomed. Biotechnol., 2012:484869.

[32]Sun, E.L., Wei, W.J., Wang, L.N., et al., 2015. Musashi-1 maintains blood–testis barrier structure during spermatogenesis and regulates stress granule formation upon heat stress. Mol. Biol. Cell, 26(10):1947-1956.

[33]Wang, W., Liang, D.J., Song, X.J., et al., 2015. Magnolol inhibits the inflammatory response in mouse mammary epithelial cells and a mouse mastitis model. Inflammation, 38(1):16-26.

[34]Wu, C.L., Wang, H.Y., Xu, J., et al., 2014. Magnolol inhibits tumor necrosis factor-α-induced ICAM-1 expression via suppressing NF-κB and MAPK signaling pathways in human lung epithelial cells. Inflammation, 37(6):1957-1967.

[35]Wu, J., Wu, S.H., Bollig, A., et al., 2009. Identification of the cyclin D1b mRNA variant in mouse. Mol. Biol. Rep., 36(5):953-957.

[36]Xia, X., Wang, H., Niu, X., et al., 2014. Assessment of the anti-diarrhea function of compound Chinese herbal medicine cangpo oral liquid. Afr. J. Tradit. Complement. Altern. Med., 11(1):140-147.

[37]Yan, J., Liu, X.L., Han, L.Z., et al., 2015. Relation between Ki-67, ER, PR, Her2/neu, p21, EGFR, and TOP II-α expression in invasive ductal breast cancer patients and correlations with prognosis. Asian Pac. J. Cancer Prev., 16(2):823-829.

[38]Yin, J., Liu, M., Ren, W., et al., 2015a. Effects of dietary supplementation with glutamate and aspartate on diquat-induced oxidative stress in piglets. PLoS ONE, 10(4):e0122893.

[39]Yin, J., Ren, W., Yang, G., et al., 2015b. L-Cysteine metabolism and its nutritional implications. Mol. Nutr. Food Res., 60(1):134-146.

[40]Yin, J., Wu, M., Duan, J., et al., 2015c. Pyrrolidine dithiocarbamate inhibits NF-κB activation and upregulates the expression of Gpx1, Gpx4, occludin, and ZO-1 in DSS-induced colitis. Appl. Biochem. Biotechnol., 177(8):1716-1728.

[41]Yu, J., Yin, P., Yin, J., et al., 2010. Involvement of ERK1/2 signalling and growth-related molecules’ expression in response to heat stress-induced damage in rat jejunum and IEC-6 cells. Int. J. Hyperther., 26(6):538-555.

[42]Zhang, M.H., Shi, Z.D., Yu, J.C., et al., 2015. Scrotal heat stress causes sperm chromatin damage and cysteinyl aspartate-spicific proteinases 3 changes in fertile men. J. Assist. Reprod. Genet., 32(5):747-755.

[43]Zhang, Y., Zhou, Z.W., Jin, H., et al., 2015. Schisandrin B inhibits cell growth and induces cellular apoptosis and autophagy in mouse hepatocytes and macrophages: implications for its hepatotoxicity. Drug Des. Dev. Ther., 9:2001-2027.

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